Methods for Modulating the Pathway of NF-κB Using Short Hairpin RNA (ShRNA)

Marta Moretti; Barbara Di Francesco; Mauro Di Vito Nolfi; Annapaola Angrisani; Enrico De Smaele

doi:10.1007/978-1-0716-1669-7_6

Methods for Modulating the Pathway of NF-κB Using Short Hairpin RNA (ShRNA)

Methods Mol Biol. 2021:2366:95-107. doi: 10.1007/978-1-0716-1669-7_6.

Authors

Marta Moretti¹, Barbara Di Francesco², Mauro Di Vito Nolfi², Annapaola Angrisani³, Enrico De Smaele⁴

Affiliations

¹ Department of Experimental Medicine, Sapienza University of Rome, Rome, Italy.
² Department of Biotechnological and Applied Clinical Sciences, University of L'Aquila, L'Aquila, Italy.
³ Department of Molecular Medicine, Sapienza University of Rome, Rome, Italy.
⁴ Department of Experimental Medicine, Sapienza University of Rome, Rome, Italy. enrico.desmaele@uniroma1.it.

PMID: 34236634
DOI: 10.1007/978-1-0716-1669-7_6

Abstract

Target gene silencing is a strategy that can be used to turn off pathways or genes which are difficult to turn off pharmacologically, both because of lack of targeting drugs, or because of the risk of wider off-target effects. Here we describe the design and use of short hairpin RNA (ShRNA) and lentiviral vectors as an efficient technique for silencing NF-kappaB (NF-κB) pathway in cultured cells. This method can be used also in hard to transfect primary cell cultures.

Keywords: Gene silencing; Lentiviral vectors; NF-kB; RelA; ShRNA; Transduction.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Gene Silencing
Genetic Vectors / genetics
Lentivirus / genetics
Lentivirus / metabolism
NF-kappa B / genetics
NF-kappa B / metabolism
RNA Interference
RNA, Small Interfering / genetics*

Substances

NF-kappa B
RNA, Small Interfering